In recent studies, the glutamate (Glu) level has been quantified using the modified STEAM sequence on 3T MRI. We enrolled 15 healthy volunteers and a group of 51 patients who experienced stroke for the first time and had a good prognosis. The patients with infarction were divided into three groups according to their scores by using the DSM-IV diagnostic criteria for major depressive disorder and the 17-item Hamilton Depression Rating Scale (HDRS). We studied the association between post-stroke depression and (1)H-MRS measurements in unaffected frontal lobes. Single-voxel proton magnetic resonance spectroscopy ((1)H-MRS) was performed to assess N-acetylaspartate/creatine (NAA)/Cr, (Glu)/Cr, choline (Cho)/Cr, and myoinositol (mI)/Cr ratios in stroke patients. The 11 patients (21.5%) who met the criteria for depression and 9 patients (17.6%) who had a high score for HDRS, (>14) but were not depressed, had a significantly higher Glu/Cr ratio than patients who scored ≤14 on HDRS and control groups (p < 0.001). No differences were found in NAA/Cr, Cho/Cr, or mI/Cr between the groups after stroke. These findings suggest that post-stroke depression is accompanied by changes in glutamate levels in the frontal lobe.